Michelle S. Teng et al. (2006) European Worm Meeting "Expression of Mammalian GPCRs in C. elegans Generates Novel Behavioural Responses to Human Ligands"

John McCafferty, Martijn P.J. Dekkers, Andrew G. Fraser, Michelle S. Teng, Bee Ling Ng, Gert Jansen, Suzanne Rademakers

[European Worm Meeting, 2006]

Michelle S. Teng1, Martijn P.J. Dekkers2, Bee Ling Ng1, Suzanne Rademakers2, Gert Jansen2, Andrew G. Fraser1 & John McCafferty1. G protein coupled receptors (GPCRs) play a crucial role in many biological processes and represent a major class of drug targets. However purification of GPCRs for biochemical study is difficult and most methods of screening receptor-ligand interactions require cultured cells and endotoxin free compounds. In contrast, Caenorhabditis elegans is a soil dwelling nematode that feeds on bacteria and uses GPCRs expressed in chemosensory neurons to detect bacteria and environmental compounds. Here we report that expression of the mammalian somatostatin receptor (Sstr2) and chemokine receptor 5 (CCR5) in gustatory neurons allow C. elegans to specifically detect and respond to human somatostatin and MIP-1? respectively in a simple avoidance assay. The endogenous signalling components involved in this remarkable promiscuity of interaction, spanning 800 million years of evolution, are investigated. This system has practical utility in ligand screening. Using structure:function studies, we identified key amino acid residues involved in the interaction of somatostatin with its receptor. This in vivo system, which imparts novel avoidance behaviour on C. elegans, can therefore be used in screening impure GPCR ligands, including the identification of bacterial clones expressing agonists within recombinant libraries.

Barros YVR et al. (2024) Anticancer Agents Med Chem "Bee Venom Toxic Effect on MDA-MB-231 Breast Cancer Cells and Caenorhabditis Elegans."

Gubert P, da Silva LPD, Nogueira MCBL, Barros YVR, Moura AV, Porcari AM, Alves LC, Bezerra IC, Mousinho KC, Pedroza LAL, Rosini Silva AA, de Andrade AO, Antoniolli AR, Lima Filho JL, Cavalcanti IDL

[Anticancer Agents Med Chem, 2024]

INTRODUCTION: Bee venom has therapeutics and pharmacological properties. Further toxicological studies on animal models are necessary due to the severe allergic reactions caused by this product. METHOD: Here, Caenorhabditis elegans was used as an in vivo toxicity model, while breast cancer cells were used to evaluate the pharmacological benefits. The bee venom utilized in this research was collected from Apis mellifera species found in Northeast Brazil. The cytotoxicity caused by bee venom was measured by MTT assay on MDA-MB-231 and J774 A.1 cells during 24 - 72 hours of exposure. C. elegans at the L4 larval stage were exposed for three hours to M9 buffer or bee venom. Survival, behavioral parameters, reproduction, DAF-16 transcription factor translocation, the expression of superoxide dismutase (SOD), and metabolomics were analyzed. Bee venom suppressed the growth of MDA-MB-231 cancer cells and exhibited cytotoxic effects on macrophages. Also, decreased C. elegans survival impacted its behaviors by decreasing C. elegans feeding behavior, movement, and reproduction. RESULTS: Bee venom did not increase the expression of SOD-3, but it enhanced DAF-16 translocation from the cytoplasm to the nucleus. C. elegans metabolites differed after bee venom exposure, primarily related to aminoacyl- tRNA biosynthesis, glycine, serine and threonine metabolism, and sphingolipid and purine metabolic pathways. Our findings indicate that exposure to bee venom resulted in harmful effects on the cells and animal models examined. CONCLUSION: Thus, due to its potential toxic effect and induction of allergic reactions, using bee venom as a therapeutic approach has been limited. The development of controlled-release drug strategies to improve this natural product's efficacy and safety should be intensified.

Gubert, Priscila et al. (2021) International Worm Meeting "Honey bee (Apis mellifera) venom toxicity in breast cancer cells and the nematode Caenorhabditis elegans"

Aleixo Leal, Lucas, Ribeiro Barros, Yaskara Veruska, Pereira Dantas da Silva, Larissa, Cerqueira Mousinho, Kristiana, Gubert, Priscila, Roberto Antoniolli, Angelo, Onduras de Andrade, Amanda, Dillion Lima Cavalcanti, Iago, Conrado Bezerra, Iverson, Cajuba de Britto Lira Nogueira, Mariane

[International Worm Meeting, 2021]

Bee venom, also known as apitoxin, is produced by honey bees (Apis mellifera) and comprises a complex mixture of substances with reported therapeutics and pharmacological properties. However, this natural product can cause severe allergic reactions, and further toxicological studies on animal models are necessary to ensure safe use. Bee venom obtained from Apis mellifera and collected in Northeast Brazil was used to investigate its effects in normal and breast cancer cells and the nematode Caenorhabditis elegans. In the present study, we employed the acute exposure assay system of C. elegans to evaluate bee venom's toxicity in vivo. Synchronized L4 larval stage worms (N2-Bristol) were exposed for three hours in M9 buffer to bee venom. Behavioral parameters, including reproduction, survival, DAF-16 transcription factor location (zIs356 [daf-16p::daf-16a/b::GFP + rol-6(su1006)]), and superoxide dismutase-3 (SOD-3; muIs84 [(pAD76) sod-3p::GFP + rol-6(su1006)]) expression, were analyzed. Bee venom cytotoxic impacts on MDA-MB-231 and J774 A.1 cells were evaluated by the MTT assay until 72 hours of exposure. Acute exposure to bee venom resulted in a decrease in C. elegans survival, feeding behavior (p<0.0001), movement (p<0.0001) while induced an increase in the gaps between the cycles of defecation (p<0.001). Bee venom has also decreased nematode reproduction by reducing both egg-production (p<0.0001) and egg-laying (p<0.05). This toxin enhanced DAF-16 translocation from the cytoplasm to the nucleus, which did not affect the SOD-3 expression. Bee venom significantly inhibited the proliferation of MDA-MB-231 cells and caused a cytotoxic effect on macrophages. Our results show that exposure to bee venom produced significant toxic effects on the cells and animal model studied. C. elegans can provide information about the molecular and cellular mechanisms of bee venom toxicity and serve as a model organism to study the toxic effects of this natural product on human health.

McAfee A et al. (2015) Data Brief ""Controlled, cross-species dataset for exploring biases in genome annotation and modification profiles"."

Foster LJ, Michaud S, McAfee A

[Data Brief, 2015]

Since the sequencing of the honey bee genome, proteomics by mass spectrometry has become increasingly popular for biological analyses of this insect; but we have observed that the number of honey bee protein identifications is consistently low compared to other organisms [1]. In this dataset, we use nanoelectrospray ionization-coupled liquid chromatography-tandem mass spectrometry (nLC-MS/MS) to systematically investigate the root cause of low honey bee proteome coverage. To this end, we present here data from three key experiments: a controlled, cross-species analyses of samples from Apis mellifera, Drosophila melanogaster, Caenorhabditis elegans, Saccharomyces cerevisiae, Mus musculus and Homo sapiens; a proteomic analysis of an individual honey bee whose genome was also sequenced; and a cross-tissue honey bee proteome comparison. The cross-species dataset was interrogated to determine relative proteome coverages between species, and the other two datasets were used to search for polymorphic sequences and to compare protein cleavage profiles, respectively.

Michelle S Teng et al. (2005) International Worm Meeting "Reprogramming olfactory behavioural response in C elegans"

Andy Fraser, John McCafferty, Nigel Carter, Bee Ling Ng, Michelle S Teng, Gert Jansen

[International Worm Meeting, 2005]

Odour detection in animals is achieved by virtue of a large family of Olfactory Receptors (OR) expressed in the olfactory epithelium. In humans, the repertoire of odorant receptor genes consists of 1000 genes, each of which encodes a G Protein Coupled Receptor or GPCR. About half of human ORs are pseudogenes. C elegans has 1000 putative OR genes, of which 30% are pseudogenes. Functional expression of ORs in C elegans provides a valuable and novel research tool to efficiently screen for olfactory ligands as well as to study receptor ligand interaction at mid-throughput using chemotaxis response as a quantitative measure. Rat I7 has been functionally expressed in AWA and AWB neurons, which generate attraction and avoidance responses respectively (Milani et al. 2002), resulting in a response to an altered response to octanal and other ligands. We have replicated and extended the initial published observations using a wider range of ligands. Using MoFlo, we have shown that it is possible to separate live transgenic worms co-expressing elt-2 GFP marker from the non-expressors as well as sort them at various stages of the life cycle based on size prior to using the adult animals for chemotaxis assay. We have also extended AWA/AWB targeted expression to another mammalian OR- hOR17-4, which has previously shown to be expressed in both olfactory epithelium and human spermatozoa where it appears to play a role in chemosensory signalling pathways and sperm chemotaxis (Parmentier et al. 1992; Vanderhaeghen et al. 1993). hOR17-4 has been shown to respond to bourgeonal and lilial in HEK293 cells expressing hOR17-4 and in in vitro sperm chemotaxis assays. We were able to reproduce this observation in a simple chemotaxis assay by expressing hOR17-4 driven by odr-10 promoter that targets expression in the AWA neuron. Our preliminary results show a dose dependent migration towards bourgeonal in these transgenic strains at similar concentrations to that published by Spehr et al. (2004).

Michelle Teng et al. (2007) International Worm Meeting "Expression of mammalian GPCRs in C. elegans generates novel behavioural responses to human ligands."

Martijn Dekkers, Michelle Teng, Bee Ling Ng, Suzanne Rademakers, Andy Fraser, Gert Jansen, John McCafferty

[International Worm Meeting, 2007]

G-protein-coupled receptors (GPCRs) play a crucial role in many biological processes and represent a major class of drug targets. However, purification of GPCRs for biochemical study is difficult and current methods of studying receptor-ligand interactions involve in vitro systems. Caenorhabditis elegans is a soil-dwelling, bacteria-feeding nematode that uses GPCRs expressed in chemosensory neurons to detect bacteria and environmental compounds, making this an ideal system for studying in vivo GPCR-ligand interactions. We sought to test this by functionally expressing two medically important mammalian GPCRs, somatostatin receptor 2 (Sstr2) and chemokine receptor 5 (CCR5) in the gustatory neurons of C. elegans. Expression of Sstr2 and CCR5 in gustatory neurons allow C. elegans to specifically detect and respond to somatostatin and MIP-1alpha respectively in a robust avoidance assay. We demonstrate that mammalian heterologous GPCRs can signal via different endogenous G subunits in C. elegans, depending on which cells it is expressed in. Furthermore, pre-exposure of GPCR transgenic animals to its ligand leads to receptor desensitisation and behavioural adaptation to subsequent ligand exposure, providing further evidence of integration of the mammalian GPCRs into the C. elegans sensory signalling machinery. In structure-function studies using a panel of somatostatin-14 analogues, we identified key residues involved in the interaction of somatostatin-14 with Sstr2. Our results illustrate a remarkable evolutionary plasticity in interactions between mammalian GPCRs and C. elegans signalling machinery, spanning 800 million years of evolution. This in vivo system, which imparts novel avoidance behaviour on C. elegans, thus provides a simple means of studying and screening interaction of GPCRs with extracellular agonists, antagonists and intracellular binding partners.

Munch D et al. (2008) Funct Ecol "Ageing in a eusocial insect: molecular and physiological characteristics of life span plasticity in the honey bee."

Wolschin F, Amdam GV, Munch D

[Funct Ecol, 2008]

Commonly held views assume that ageing, or senescence, represents an inevitable, passive, and random decline in function that is strongly linked to chronological age. In recent years, genetic intervention of life span regulating pathways, for example, in Drosophila as well as case studies in non-classical animal models, have provided compelling evidence to challenge these views.Rather than comprehensively revisiting studies on the established genetic model systems of ageing, we here focus on an alternative model organism with a wild type (unselected genotype) characterized by a unique diversity in longevity - the honey bee.Honey bee (Apis mellifera) life span varies from a few weeks to more than 2 years. This plasticity is largely controlled by environmental factors. Thereby, although individuals are closely related genetically, distinct life histories can emerge as a function of social environmental change.Another remarkable feature of the honey bee is the occurrence of reverted behavioural ontogeny in the worker (female helper) caste. This behavioural peculiarity is associated with alterations in somatic maintenance functions that are indicative of reverted senescence. Thus, although intraspecific variation in organismal life span is not uncommon, the honey bee holds great promise for gaining insights into regulatory pathways that can shape the time-course of ageing by delaying, halting or even reversing processes of senescence. These aspects provide the setting of our review.We will highlight comparative findings from Drosophila melanogaster and Caenorhabditis elegans in particular, and focus on knowledge spanning from molecular- to behavioural-senescence to elucidate how the honey bee can contribute to novel insights into regulatory mechanisms that underlie plasticity and robustness or irreversibility in ageing.

Zain MM et al. (2016) Trop Life Sci Res "The Effect of Macrocyclic Lactones-Ivermectin Exposure on Egg Hatching and Larval Development of Caenorhabditis elegans."

Zain MM, Him NA, Yahaya ZS

[Trop Life Sci Res, 2016]

To date, the ivermectin resistance in nematode parasites has been reported and many studies are carried out to determine the causes of this problem. A free-living Caenorhabditis elegans is used as a model system for this study to investigate the response of C. elegans to ivermectin exposure by using larval development assay. Worms were exposed to ivermectin at concentration from 1 ng/mL to 10 ng/mL and dimethyl sulphoxide (DMSO) as a control. The developments of the worms were monitored for 24, 48, 72, and 96 hours until the worms become adults. Results indicated that worms' growth began to be affected by ivermectin at a concentration of 5 ng/mL, while at the concentration of 6, 7, 8, 9, and 10 ng/mL, the growth of worms were inhibited compared to control worms. Further study of the protein expression in C. elegans should be done to investigate the up-regulated and down-regulated proteins involve in ivermectin resistance.

Hussain R et al. (1981) J Immunol "IgE responses in human filariasis. I. Quantitation of filaria-specific IgE."

Ottesen EA, Hussain R, Hamilton RG, Adkinson NF, Kumaraswami V

[J Immunol, 1981]

We have developed a noncompetitive solid phase radioimmunoassay to quantitate human IgE antibodies against soluble adult antigens of Brugia malayi (B.m.), a filarial parasite, in sera of patients with various forms of clinical filariasis in Madras, India. A single reference serum was shown to contain 23 micrograms/ml of B.m.-specific IgE by depletion analysis and was used as a standard serum throughout the study. The levels of specific IgE ranged in the patients sera from 2 to 23,000 ng/ml. When these individuals were divided into clinical groups, the individuals with tropical pulmonary eosinophilia had the highest levels (mean = 8630 ng/ml) and were significantly higher than all the other groups (p less than 0.001). The lowest levels were seen in patients with circulating microfilariae (mean = 30.5 ng/ml). Patients exhibiting lymphatic obstruction (i.e., chronic pathology group) had levels slightly higher than microfilaremics (mean = 68 ng/ml) but were not significantly different (p less than 0.1). Surprisingly, individuals living in endemic areas but who had no clinical signs of filariasis also showed appreciable levels of B.m.-specific IgE (mean = 55 ng/ml). The B.m.-specific IgE represented 0.1 to 48% of the total IgE. High percentages of specific IgE may be responsible for evoking allergic symptomatology in patients with tropical pulmonary eosinophilia.

Haldimann P et al. (2011) J Biol Chem "The novel hydroxylamine derivative NG-094 suppresses polyglutamine protein toxicity in Caenorhabditis elegans."

Goloubinoff P, Muriset M, Vigh L, Haldimann P

[J Biol Chem, 2011]

Aggregation-prone polyglutamine (polyQ) expansion proteins cause several neurodegenerative disorders, including Huntington disease. The pharmacological activation of cellular stress responses could be a new strategy to combat protein conformational diseases. Hydroxylamine derivatives act as co-inducers of heat-shock proteins (HSPs) and can enhance HSP expression in diseased cells, without significant adverse effects. Here, we used Caenorhabditis elegans expressing polyQ expansions with 35 glutamines fused to the yellow fluorescent protein (Q35-YFP) in body wall muscle cells as a model system to investigate the effects of treatment with a novel hydroxylamine derivative, NG-094, on the progression of polyQ diseases. NG-094 significantly ameliorated polyQ-mediated animal paralysis, reduced the number of Q35-YFP aggregates and delayed polyQ-dependent acceleration of aging. Micromolar concentrations of NG-094 in animal tissues with only marginal effects on the nematode fitness sufficed to confer protection against polyQ proteotoxicity, even when the drug was administered after disease onset. NG-094 did not reduce insulin/insulin-like growth factor 1-like signaling, but conferred cytoprotection by a mechanism involving the heat-shock transcription factor HSF-1 that potentiated the expression of stress-inducible HSPs. NG-094 is thus a promising candidate for tests on mammalian models of polyQ and other protein conformational diseases.